LOCUS       sequence-66-expo        3633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3633)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 3633)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pPD79_44
FEATURES             Location/Qualifiers
     source          1..3633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..35
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     19..35
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             31..1278
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPVEKMSKGEELFTGVVPILVELDGD
                     VNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKR
                     HDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGH
                     KLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDN
                     HYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKCPGDPLVLERPPPRWSSNS
                     PYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLN
                     GEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAGFPRQALNRGLPL
                     GFRFSALRHLDPKKLD"
     primer_bind     54..74
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        56..74
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             103..816
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(133..161)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     756..776
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(826..842)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(826..842)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(874..893)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(875..893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(903..920)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(903..919)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(912..934)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1061..1516
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1148..1167)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1358..1379
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1542..1646
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1647..1715,1716..2507)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1865..1884)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2678..3266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3167..3186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3420..3437
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3554..3575
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3590..3595,3596..3613,3614..3620)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    join(3628..3633,1..11)
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     join(3633,1..22)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
       61 accctcacta aagggaacaa aagctgggta ccggtagaaa aaatgagtaa aggagaagaa
      121 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
      181 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
      241 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttctcttat
      301 ggtgttcaat gcttttcaag atacccagat catatgaaac ggcatgactt tttcaagagt
      361 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
      421 aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
      481 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
      541 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa
      601 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
      661 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
      721 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
      781 gctgggatta cacatggcat ggatgaacta tacaaatgcc cgggggatcc actagttcta
      841 gagcggccgc caccgcggtg gagctccaat tcgccctata gtgagtcgta ttacgcgcgc
      901 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
      961 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     1021 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     1081 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1141 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1201 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1261 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1321 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1381 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1441 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1501 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1561 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1621 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1681 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1741 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1801 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1861 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1921 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1981 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2041 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2101 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2161 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2221 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2281 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2341 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2401 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2461 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2521 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2821 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3301 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3361 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3421 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3481 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3541 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     3601 tgcttccggc tcgtatgttg tgtggaattg tga
//
